摘要
目的研究7-二氟亚甲基-5,4'-二甲氧基染料木黄酮对H_2O_2诱导血管内皮细胞凋亡的影响,并初步探讨其作用机制。方法用1 mmol/L H_2O_2诱导人脐静脉内皮细胞建立氧化应激损伤模型。实验分为空白对照组、H_2O_2损伤组、溶媒对照组、先导化合物组(100μmol/L染料木黄酮)、不同浓度(0.1、0.3、1、3、10μmol/L)7-二氟亚甲基-5,4'-二甲氧基染料木黄酮组。DCFH-DA激活荧光流式细胞术测定活性氧的生成,吖啶橙染色荧光显微镜观察细胞凋亡形态,PI染色流式细胞术检测细胞凋亡率,Western blot测定Caspase-3的表达。结果 1 mmol/L H_2O_2孵育血管内皮细胞24 h,活性氧的生成显著增加,吖啶橙染色荧光显微镜下细胞呈现明显的凋亡形态学改变,细胞凋亡率升高,Caspase-3的表达上调。用7-二氟亚甲基-5,4'-二甲氧基染料木黄酮预处理血管内皮细胞后可见活性氧的生成减少,细胞凋亡率降低,Caspase-3的表达下调。结论 7-二氟亚甲基-5,4'-二甲氧基染料木黄酮对H_2O_2诱导血管内皮细胞凋亡有明显的抑制作用,其作用机制可能是通过降低活性氧的生成,下调Caspase-3的表达,从而拮抗H_2O_2诱导的血管内皮细胞凋亡。
Aim To study the effects of 7-difluoromethyl-5,4＇-dimethoxygenistein（d FMGEN） on the apoptosis to H2O2-induced vascular endothelial cells and its molecular mechanism. Methods The oxidative stress injury model was established by human umbilical vein endothelial cells induced by 1 mmol/L H2O2. The experiment was divided into blank control group,H2O2 group,vehicle group,100 μmol/L genistein group and different concentrations（0.1,0.3,1,3,10 μmol/L） d FMGEN groups. The reactive oxygen species（ROS） generation was examined by DCFH-DA activated fluorescence flow cytometry（FCM）. Apoptotic morphology was observed by acridine orange staining under fluorescence microscopy. The apoptotic rate was detected by PI staining FCM. The expression of Caspase-3 was determined by Western blot. Results When vascular endothelial cells were treated with 1 mmol/L H2O2 for 24 h,the generation of ROS was highly added,obvious morphological changes of apoptosis was found by acridine orange staining under fluorescence microscopy,the apoptotic rate was increased and the expression of Caspase-3 was up-regulated. After pretreating with d FMGEN,results showed that d FMGEN could reduce the release of ROS,decrease the apoptotic rate and down-regulate the expression of Caspase-3. Conclusion d FMGEN can effectually inhibit the apoptosis of vascular endothelial cells induced by H2O2,which is probably due to the decrease of ROS generation and downregulation of Caspase-3,thereby inhibiting apoptosis of vascular endothelial cells induced by H2O2.
出处
《中国动脉硬化杂志》
CAS
北大核心
2017年第12期1219-1224,共6页
Chinese Journal of Arteriosclerosis
基金
湖南省教育厅科研资助项目(10C1178)
